Effects of a patient-derived de novo coding alteration of CACNA1I in mice connect a schizophrenia risk gene with sleep spindle deficits

摘要

The exon/intron structure of mouse , and the positions of targeting sgRNAs for generating the Ca 3.3 knock-out (KO) and R1305H knock-in animals. The sequences of the targeting sgRNAs and the genomic sequence verification for knock-out and knock-in founder animals are shown underneath. Western blot from and whole-brain lysates showing a near complete lack of Ca 3.3 protein in the (−/−) samples. Significant reductions of both mRNA and protein levels in the heterozygous KO mice were also found (Supplementary Fig. ). Bottom panel: quantification of Ca 3.3 protein and mRNA levels normalized to levels (  = 5). A typical representative western blot showing total Ca 3.3 protein levels (top panel) and Ca 3.3 protein levels in crude synaptoneurosomal preparations (bottom panel) from brain tissues derived from littermates of (+/+) and (RH/RH). Quantification of total and synaptoneurosomal Ca 3.3 levels in the total brain lysate (  = 8) and crude synaptoneurosomal preparation (  = 6). A typical western blot showing the developmental progression of Ca 3.3 expression in brain tissue across different postnatal days, and at 5 (5m) and 8 months (8m). Quantification of protein (  = 2 for each time point) and mRNA (  = 2 for each time point except for 5m and 8m that contains  = 3 each) levels during developmental progression. **